OBJECTIVE: Benzylisoquinoline alkaloids (BIAs) have pharmacological functions and clinical use. BIAs are mainly distributed in plant species across the order Ranunculales and the genus Phellodendron from Sapindales. The BIA biosynthesis has been intensively investigated in Ranunculales species. However, the accumulation mechanism of BIAs in Phellodendron is largely unknown. The aim of this study is to unravel the biosynthetic pathways of BIAs in Phellodendron amurens. METHODS: The transcriptome and metabolome data from 18 different tissues of P. amurense were meticulously sequenced and subsequently subjected to a thorough analysis. Weighted gene co-expression network analysis (WGCNA), a powerful systems biology approach that facilitates the construction and subsequent analysis of co-expression networks, was utilized to identify candidate genes involved in BIAs biosynthesis. Following this, recombinant plasmids containing candidate genes were expressed in Escherichia coli, a widely used prokaryotic expression system. The purpose of this genetic engineering endeavor was to express the candidate genes within the bacteria, thereby enabling the assessment of the resultant enzyme activity. RESULTS: The synonymous substitutions per synonymous site for paralogs indicated that at least one whole genome duplication event has occurred. The potential BIA biosynthetic pathway of P. amurense was proposed, and two PR10/Bet v1 members, 14 CYP450s, and 33 methyltransferases were selected as related to BIA biosynthesis. One PR10/Bet v1 was identified as norcoclaurine synthase, which could catalyze dopamine and 4-hydroxyphenylacetaldehyde into (S)-norcoclaurine. CONCLUSION Our studies provide important insights into the biosynthesis and evolution of BIAs in non-Ranunculales species.

Objective To compare the effectiveness of femoral neck system(FNS)and inverted triangle cannulated screws(ITCS)in the treatment of femoral neck fractures in young adults.Methods A retrospective study was conducted on 106 young and middle-aged patients who had been surgically treated for femoral neck fracture at our hospital from December 2020 to June 2022.The patients were assigned to FNS group(57 cases)or ITCS group(49 cases)according to the different internal fixations.The operation time,intraoperative fluoroscopy frequency,intraoperative blood loss,extra assisted reduction procedures(Kirschner wire prying reduction or open reduction),hospital stays,healing time,complications,postoperative recovery,and follow-up duration were compared between the two groups.Results The operation time,fluoroscopy frequency,and healing time in the FNS group were less than those in the ITCS group(P<0.001).The shortening degree and length of femoral neck in moderate and severe patients of the FNS group were lower than those of the ITCS group(P<0.001).The Harris score of the FNS group was higher than that of the ITCS group 12 months after surgery(P<0.001).There was no significant difference in intraoperative blood loss,reduction assistance,Garden index,or hospital stays between the two groups(P>0.05).Conclusion FNS is a better choice for internal fixation in patients with femoral neck fractures than ITCS,with faster recovery and less postoperative complications.

Objective To compare the effectiveness of femoral neck system(FNS)and inverted triangle cannulated screws(ITCS)in the treatment of femoral neck fractures in young adults.Methods A retrospective study was conducted on 106 young and middle-aged patients who had been surgically treated for femoral neck fracture at our hospital from December 2020 to June 2022.The patients were assigned to FNS group(57 cases)or ITCS group(49 cases)according to the different internal fixations.The operation time,intraoperative fluoroscopy frequency,intraoperative blood loss,extra assisted reduction procedures(Kirschner wire prying reduction or open reduction),hospital stays,healing time,complications,postoperative recovery,and follow-up duration were compared between the two groups.Results The operation time,fluoroscopy frequency,and healing time in the FNS group were less than those in the ITCS group(P<0.001).The shortening degree and length of femoral neck in moderate and severe patients of the FNS group were lower than those of the ITCS group(P<0.001).The Harris score of the FNS group was higher than that of the ITCS group 12 months after surgery(P<0.001).There was no significant difference in intraoperative blood loss,reduction assistance,Garden index,or hospital stays between the two groups(P>0.05).Conclusion FNS is a better choice for internal fixation in patients with femoral neck fractures than ITCS,with faster recovery and less postoperative complications.

Background Accurate T-staging is critical for esophageal cancer therapy,but CT-based assessment has significant limitations.Large vision models(LVMs)hold promise,yet their zero-shot clinical diagnostic capability without fine-tuning remains unvalidated.Methods A retrospective analysis was conducted on the chest CT images from 98 esophageal cancer patients and 50 normal controls.Using radiologist-consensus as the gold standard,the zero-shot T-staging performance of 3 LVMs(GPT-5,Gemini,and MedGemma)was evaluated with prompts of varying complexity.Results GPT-5 exhibited the highest accuracy and stability.Significant biases were observed among models:Gemini tended to over-stage,while MedGemma showed a tendency to under-stage.All models faced challenges in identifying early-stage tumors,but structured prompts improved diagnostic performance for mid-to-late stage lesions.Conclusion LVMs have potential for zero-shot T-staging,but their performance highly depends on model choice and prompt design.The generic model GPT-5 show superior zero-shot generalization.However,current model performance is not yet clinically viable,especially for early diagnosis.Future work should focus on fine-tuning with high-quality clinical data and developing standardized prompt frameworks.

Background Accurate T-staging is critical for esophageal cancer therapy,but CT-based assessment has significant limitations.Large vision models(LVMs)hold promise,yet their zero-shot clinical diagnostic capability without fine-tuning remains unvalidated.Methods A retrospective analysis was conducted on the chest CT images from 98 esophageal cancer patients and 50 normal controls.Using radiologist-consensus as the gold standard,the zero-shot T-staging performance of 3 LVMs(GPT-5,Gemini,and MedGemma)was evaluated with prompts of varying complexity.Results GPT-5 exhibited the highest accuracy and stability.Significant biases were observed among models:Gemini tended to over-stage,while MedGemma showed a tendency to under-stage.All models faced challenges in identifying early-stage tumors,but structured prompts improved diagnostic performance for mid-to-late stage lesions.Conclusion LVMs have potential for zero-shot T-staging,but their performance highly depends on model choice and prompt design.The generic model GPT-5 show superior zero-shot generalization.However,current model performance is not yet clinically viable,especially for early diagnosis.Future work should focus on fine-tuning with high-quality clinical data and developing standardized prompt frameworks.

Objective To explore the effects of Xin Jia Congrong Tusizi decoction on mitochondrial biosynthesis and mitochondrial function in ovarian granulosa cells.Methods We prepared the functional injury model of human ovarian granulosa cells KGN induced by triptolide.We divided the cells into control group,model group,Tiao jing Cuyun Pill group,Xin Jia Congrong Tusizi decoction group,SIRT1 inhibitor group and SIRT1 inhibitor+Xin Jia Congrong Tusizi decoction group.After 6 h incubation with triptolide to cause functional impairment of granulosa cells,SIRT1 inhibitor,blank serum and medicated serum were added for 48 h.Cell viability and apoptosis rate were assessed using CCK-8 and flow cytometry.Anti-Müllerian(AMH),follicle-stimulating hormone(FSH)and inhibin B(INHB)were detected by ELISA.ATP enzyme and MMP were used to detect by biochemical kit and JC-1 method.apoptosis rate and MMP were assessed using flow cytometry.The morphology,structure,quantity and activity changes of mitochondria were observed by electron microscope and fluorescence microscope.Western blot and PCR were used to detect the protein and mRNA expressions of SIRT1,p-SIRT1,PGC-1α,NRF1 and TFAM.Results Chinese patent medicine TiaojingCuyun Pill and bushen Yangxue Huoxue therapy compound Xin Jia Congrong Tusizi decoction augmented the activity of granulosa cells(P<0.05),decreased the apoptosis rate(P<0.05),increased ATP enzyme and MMP(P<0.05),improved mitochondrial morphological and structural damage,added the mtDNA level,the number and activity of mitochondria(P<0.01)and up-regulated and the protein and mRNA expressions of SIRT1,PGC-1α,NRF1,TFAM(P<0.05).Conclusion Xin Jia Congrong Tusizi decoction protected ovarian granulosa cells damage caused by triptolide.Its mechanism may related to improve mitochondrial dysfunction by regulating SIRT1/PGC-1α signaling pathway in order to promote the biosynthesis of new mitochondria.

Objective:The treatment of head and neck in plexiform neurofibroma (PNF) is a major clinical problem, lacking consensus on surgical treatment, classification, operation timing, and treatment method. The purpose of this study was to provide a basis for further consensus formation by analyzing the clinical manifestations, surgical conditions, tumor recurrence, post-operation satisfaction, and changes in quality of life of patients undergoing PNF surgery in head and neck.Methods:Through medical record review and telephone follow-up, a retrospective analysis was conducted on neurofibromatosis type 1 (NF1) patients admitted for surgical treatment for PNF patient in head and neck from May 2012 to July 2022 in Department of Plastic and Reconstructive Surgery, Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine. Complete collection and statistical analysis of patients’clinical data, using telephone follow-up to investigate the immediate postoperative satisfaction and long-term surgical satisfaction of patients and/or their families, as well as standardized quality of life questionnaires HRQol(health related quality of life) and PlexiQol(plexiform neurofibroma quality of life). Based on the data about changes in quality of life before and after surgery and long-term surgical satisfaction, patients were divided into surgical benefit and non-benefit groups. Binary and multivariate logistic regression analysis were used to analyze the clinical characteristics of patients with long-term surgical benefit.Results:Totally 512 patients with head and neck NF1 were admitted for surgery with complete medical records. 121 patients were identified as NF1 related PNF diagnosed by medical history and radiological examination, and effective follow-up was obtained. There were 70 males and 51 females, aged (25.60±12.85) years old, ranging from 7 to 63 years old, with 41 patients who were ≤ 18 years old and 80 patients over 18 years old. 62.81%(76/121) of patients exhibiting clinical dysfunctions, and the tumor mass were mainly characterized by invasive growth. 41.32%(50/121) of patients underwent multiple surgical treatments, with a total of 215 surgeries performed on 121 patients. The surgical objective included appearance improvement and functional repair. The incidence of postoperative complications was 6.05%(13/215). The follow-up period after last operation was (51.41±27.66) months, and 42.15%(51/121) of patients reported postoperative tumor recurrence. 76.03%(92/121) of patients were satisfied with immediate postoperative result, while the rate decreased to 46.28%(56/121) during long-term follow-up. Family members of patients who were ≤ 18 years old had a higher proportion of dissatisfaction with the scars caused by surgery and a stronger willingness to undergo another surgery. The tumor recurrence was closely related to surgical benefits ( OR=2.32, P<0.05). Further analysis found that the gender and age of patients were the main risk factors for the recurrence. The recurrence risk in patients ≤ 18 years old was significantly higher than in that over 18 years old( OR=3.49, P=0.004), and the highest in the 7-12 year-old group, reaching 68.42%(13/19). The recurrence risk in male patients was significantly lower than that in females ( OR=0.40, P=0.026). Conclusion:The clinical manifestations of PNF patients in head and neck region are complex. Clinical diagnosis and treatment in PNF should focus on the applications in comprehensive method such as full preoperative evaluation, active multi-disciplinary treatment cooperation and combined therapies in order to improve the safety and effectiveness of treatment and reduce tumor recurrence.

Objective To explore the impact of high glucose(HG)intervention on immune escape of pancreatic cancer cells and its molecular mechanisms.Methods PANC-1 cells were treated with different concentrations of glucose(0,7.5,15,30 mmol/L)for 24 h to establish high glucose intervention PANC-1 cells.miR-429 mimics and its negative control(mimics NC)were transfected into PANC-1 cells,which were divided into control group,HG group,HG+mimics NC group,HG+mimics group,HG+mimics+oe-NC group,and HG+mimics+oe-ZEB1 group.Flow cytometry was utilized to measure the expression level of cell surface molecule PD-L1;qRT-PCR was used to detect the expression levels of miR-429 and ZEB1 mRNA in cells;Western blot was used to detect the ex-pression level of ZEB1 protein in cells.The above-mentioned PANC-1 cells from each group were co-cultured with CD8+T cells to establish a co-culture system,and CCK-8 was used to assess cell proliferation activity;apoptosis levels of cells were measured using flow cytometry;lactate dehydrogenase(LDH)release assay was used to detect the killing effect of CD8+T cells on PANC-1 cells;dual-luciferase reporter system was used to validate the target-regulatory relationship between mi R-429 and ZEB1.Results HG could promote the expression of cell surface mole-cules PD-L1 and ZEB1 in PANC-1 cells(P＜0.05),inhibit the expression of miR-429,and exhibit concentration dependence.Overexpression of miR-429 could significantly suppress the expression of cell surface molecule PD-L1 induced by HG in PANC-1 cells,while overexpression of ZEB1 could reverse the inhibitory effect of miR-429 over-expression on the expression of cell surface molecule PD-L1 induced by HG.After establishing a co-culture system with CD8+T cells,compared with the control group,the proliferation activity of PANC-1 cells in the HG group sig-nificantly increased,and the apoptosis rate and cytotoxicity significantly decreased(P＜0.05).Compared with the HG+mimics NC group,the proliferation activity of PANC-1 cells in the HG+mimics group significantly decreased,and the apoptosis level and cytotoxicity significantly increased(P＜0.05).Compared with the HG+mimics group,the proliferation activity of PANC-1 cells in the HG+mimics+oe-ZEB1 group significantly increased,and the apop-tosis rate and cytotoxicity significantly decreased(P＜0.05).Dual luciferase reporter gene assay confirmed that miR-429 negatively regulated ZEB1.Conclusion High glucose promotes immune escape of PANC-1 cells by down-regulating the expression level of miR-429,negatively regulating the expression of ZEB1 mRNA,and increasing the expression level of cell surface molecule PD-L1 in PANC-1 cells.